This study defined the best site for quantifying osteoclasts in male Lewis rats with mycobacteria-induced adjuvant arthritis. Hind paw sections of normal and arthritic rats (n = 6 per group) taken 7 days after disease onset were stained for osteoclasts using an anti-human cathepsin K primary antibody. Erosions and osteoclasts were assessed using semiquantitative scores (entire section) and quantitative measures (in calcaneus, navicular tarsal, and tibia). Bone area in arthritic rats was significantly reduced (P </= 0.05) by 39-55%, with the greatest decrease in the tarsal. Osteoclasts in arthritic rats were significantly increased (P </= 0.05) relative to normal in calcaneus (sevenfold), tarsal (194-fold), and tibia (threefold). The most useful quantitative indices were bone area and total osteoclasts, both defined as a percentage of total area measured. Semiquantitative scores for bone erosion and osteoclasts paralleled the quantitative indices. These data show that the navicular tarsal is the most sensitive site at which to assess arthritis in rats with this variant of adjuvant-induced arthritis.

OBJECTIVETo assess the capacities of the cytokine inhibitors interleukin-1 receptor antagonist (IL-1Ra; anakinra) and PEGylated soluble tumor necrosis factor receptor I (PEG sTNFRI; pegsunercept) to suppress neovascularization.METHODSA corneal angiogenesis assay was performed by implanting nylon discs impregnated with an angiogenic stimulator (basic fibroblast growth factor or vascular endothelial growth factor) into one cornea of female Sprague-Dawley rats. Animals were treated with IL-1Ra or PEG sTNFRI for 7 days, after which new vessels were quantified. In a parallel study, male Lewis rats with mycobacteria-induced adjuvant-induced arthritis were treated with IL-1Ra or PEG sTNFRI for 7 days beginning at disease onset, after which scores for inflammation and bone erosion as well as capillary counts were acquired from sections of arthritic hind paws.RESULTSTreatment with IL-1Ra yielded a dose-dependent reduction in growth factor-induced corneal angiogenesis, while PEG sTNFRI did not. IL-1Ra, but not PEG sTNFRI, significantly reduced the number of capillaries in arthritic paws, even though both anticytokines reduced inflammation and bone erosion to a similar degree.CONCLUSIONThese data support a major role for IL-1, but not TNFalpha, in angiogenesis and suggest that an additional antiarthritic mechanism afforded by IL-1 inhibitors, but not anti-TNF agents, is the suppression of the angiogenic component of pannus.

BackgroundAdjuvant arthritis (AdA) induced in Lewis rats by injection of mycobacteria in oil is characterised by marked inflammation and skeletal damage. Recent work by our group using the AdA model has shown that skeletal destruction is mediated by the osteoclast differentiation factor osteoprotegerin (OPG) ligand,1 and that inhibition of bone loss is associated with anti-inflammatory efficacy.2ObjectivesThe current study was performed to assess whether anti-inflammatory molecules act to spare bone by regulating intra-lesional numbers of osteoclasts.MethodsAdA was induced in male Lewis rats (n = 6/group) on Day 0, and hind paw swelling developed on Day 9. Animals were treated for 7 days (Days 9–15) and then necropsied (Day 16). Therapy consisted of interleukin-1 receptor antagonist (IL-1ra) or pegylated soluble tumour necrosis factor receptor type I (PEG sTNF-RI), administered alone or in all possible combinations, at the following doses: IL-1ra, 0.2, 1.0 or 5.0 mg/kg/hr (by s.c. infusion); PEG sTNF-RI, 0.25, 1.0 or 4.0 mg/kg/day (s.c bolus). Osteoclasts were detected by an indirect immunohistochemical method for cathepsin K. Inflammation, bone erosion and osteoclast numbers were scored blindly using tiered, semi-quantitative grading scales.ResultsDose-dependent reductions in inflammation, bone erosion and osteoclast scores were observed for high doses of both anti-cytokine therapies, whether given alone or in combination.ConclusionThese data indicate that anti-inflammatory doses of IL-1ra and PEG sTNF-RI protect the structural integrity of arthritic joints by regulating osteoclast numbers.ReferencesKong, et al. Nature 1999;402:304 Feige, et al. Cell Mol Life Sci. 2000;57:1457

Background Marked polyarthritis is induced in male Lewis rats by injection of either heat-killed Mycobacterium tuberculosis in mineral oil (AdA) or lipoidal amine in complete Freund’s adjuvant (LA/CFA). For both models, paw swelling and body weight loss occur in all rats by 10 to 12 days after immunisation. Objectives Interestingly, however, the present study demonstrates that the morphologic and densitometric changes of AdA and LA/CFA are distinguishable. Methods We induced AdA or LA/CFA in male Lewis rats (180 g) and evaluated both clinical (body and spleen weight, volume of the tibiotarsal [hock] joints) and morphologic changes (bone mineral density [BMD] by DEXA; histopathologic lesions in the hock and femorotibial [knee] joints). Readings were obtained at the day of onset (as determined by initial paw swelling) as well as 5, 10 and 25 days after onset. Results Paw swelling, loss of body weight, enhanced spleen weight were identical in AdA and LA/CFA. In contrast, hock inflammation score, hock erosion score and hock cartilage score were lower in LA/CFA, though not statistically significant from AdA. Lesions developed more slowly in LA/CFA than in AdA. However, hock BMD as well as knee inflammation and erosion scores found to be different (shown here at Onset + 5 days): Conclusion Inflammatory and skeletal changes for AdA and LA/CFA were similar by clinical and histopathologic measures 10 days after onset, while inflammation receded by Onset + 25 days in LA/CFA more than it did in AdA. These data indicate that bone densitometric and morphologic findings can distinguish these two models of adjuvant arthritis despite their similar clinical presentations. This finding may be relevant in screening novel therapeutic candidates as these models may define different potential efficacious effects of a compound.

We investigated the molecular basis of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor gene transcription in immortalized alpha T3 gonadotropes, hypothalamic GT1-7 and hippocampal HN33p neurons. Nuclear run-on transcription, as well as transfection assays with fusion constructs of luciferase and the 5'-flanking region of LH/hCG receptor gene, revealed that GT1-7 neurons transcribe more than the alpha T3 or HN33p cells. Transient transfection of truncated reporter gene constructs and gel mobility shift assays revealed that while all neuroendocrine cells use the same promoter, they contain different levels of promoter binding proteins. Higher levels of these proteins may explain increased transcription of the LH/hCG receptor gene in GT1-7 neurons compared with alpha T3 and HN33p cells.

A fluorescence lifetime-based fiber-optic thermometer has been designed and characterized to carry out wide-range and real-time-temperature measurements, over a region from 77 K (liquid nitrogen) to 800 K. Computer-assisted data acquisition and digital signal-processing techniques which are used permit the taking of real-time temperature measurements. The results of the study have shown that the use of a smaller-size sensor element is preferable to avoid problems due to the redistribution of fluorescence and thus any resulting errors in the calibration, which is compared with the results of theoretical models. The extension of the work to the lower temperature range (< 200 K) highlights this problem, which has not been discussed before in temperature probe fabrication. (C) 1997 Elsevier Science S.A.

Research has been carried out showing that severe errors in the calibration of ruby fluorescence decay-time fibre optic thermometers, particularly in the cryogenic region (>77K), can occur if probes are not carefully designed to avoid fluorescence reabsorption effects. This effect is investigated and quantified

Noniterative algorithms using Prony’s method are proposed for the estimation of the exponential lifetime in fluorescence-based fiber-optic sensor systems. The uses of a variety of Prony’s algorithms are evaluated using Monte Carlo simulations. It is shown that the performance of Prony’s algorithms is quite close to that of Marquardt algorithms and they are the better alternatives to their Marquardt counterparts, when illustrated for real-time applications in temperature sensors with a 98% decrease in running time.

A condition survey was carried out on four affected concrete structures where dolomitic rocks had been used as coarse aggregates. Compositions, textures and alkali reactivities of partial coarse aggregates were determined and the deteriorated concrete cores were subjected to petrographic examination. The results revealed that the dolomitic aggregates were potentially alkali reactive and were responsible for the deterioration of the concrete structures. It was the frist time that damage to concrete by alkali-dolomite reaction was recognized in China.